PROJECT SUMMARY/ABSTRACT Transformational human health discoveries require access to advanced imaging technologies. The instruments at Purdue University's proteomics and metabolomics shared core facilities experience high usage from investigators across the state and have enabled researchers to quantify small molecules and proteins, as well as to profile the metabolome, lipidome, and proteome in biofluids and tissues. However, MS-based imaging technology has significantly advanced over the last 15 years. Therefore, the facilities' current equipment limits researchers' ability to obtain the needed quality and level of chemical information from their samples. As a result, users require a more sophisticated and powerful instrument, represented by the proposed instrument. The Waters SYNAPT G2-Si High Definition Mass Spectrometry (HDMS) system is a unique, versatile platform that provides high-resolution, ion mobility-enabled, quadrupole time-of-flight (QToF) mass spectrometry. The SYNAPT G2 is equipped with matrix-assisted laser desorption ionization (MALDI) and desorption electrospray ionization (DESI) sources that allows for both large (MALDI source) and small (DESI source) molecule imaging. The SYNAPT is the only commercially available mass spectrometry imaging (MSI) platform with MALDI or DESI imaging paired with ion mobility. Although techniques such as immunohistochemistry may achieve the same goal as MSI, these other techniques are limited in their ability to analyze multiple molecules of interest at once, and can prove to be lacking if researchers do not have prior knowledge of the chemicals being studied. MSI can analyze dozens to hundreds of biomolecules simultaneously, in general yielding both qualitative and quantitative results. Proposed applications range from mapping drug distribution and behavior across human tissues to preclinical studies that chemically identify brain cancer tissue for intraoperative devices. Advancements will be made in the areas of cancer, coronary artery disease, malaria, Parkinson's disease, pulmonary inflammation, and filoviruses. The new instrument will be used by 9 major users with 14 funded NIH-funded grants and 7 minor users with 3 funded NIH grants. The SYNAPT G2 will be placed within a core facility with an existing, well- functioning management structure with advisory committees already in place. The facility has a proven record of enabling meaningful research collaborations across multiple universities?especially through the Indiana CTSI? with a strong history of training graduate students to be independent users of its instrumentation. The diverse types of experiments that can be performed on the SYNAPT G2 system will enable investigators to define the location and amount of metabolites, lipids, peptides and proteins with high mass resolution, with the added separation of compounds based on their collision cross section (CCS). This will provide essential infrastructure to NIH-supported investigators, enabling them to accelerate their research goals and provide long-term scientific outcomes.